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Baculovirus‐driven expression and purification of glycine receptor α1 homo‐oligomers
Author(s) -
Joachim Morr,
N. Rundström,
Heinrich Betz,
Dieter Langosch,
Bertram Schmitt
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)00721-k
Subject(s) - spodoptera , autographa californica , recombinant dna , receptor , heterologous expression , sf9 , microbiology and biotechnology , membrane , biology , glycine receptor , biophysics , biochemistry , chemistry , glycine , amino acid , gene
The glycine receptor is a ligand‐gated anion channel protein of postsynaptic membranes. We expressed a homo‐oligomeric receptor composed of human α1 subunits in Spodoptera frugiperda cells by infection with a recombinant Autographa californica nuclear polyhedrosis virus. A substantial fraction of the recombinant receptor was incorporated as a functional channel protein into the cell's plasma membrane at expression levels 4‐to 30‐fold higher than in other eukaryotic heterologous expression systems or native rat spinal cord membranes, respectively. Upon detergent solubilization, the al receptor was found to exist in a predominantly monodisperse state and could be affinity‐purified to near homogeneity. This preparation is a potential starting point for future crystallisation studies.